Apparatus for extracting particle size data



May 17, 1960 G. E. GERHARDT APPARATUS FOR ExTRAcTING PARTICLE sIzE DATA med sept. 13, 195e 7 Sheets-Sheet l 1N VEN TOR. GERARD ERNEST GERHARDL' A 7' TORNE Y May 17,v 1960 G E. GERHARDT v APPARATUS FOR EXTRACTING PARTICLE SIZE DATA Filed Sept. 13. l1956 '7 Sheets-Sheet 2 SHAPED v/oEo /NPur l r DELfAl'/NE sY/vc. v sY/vc.

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l i l /f A T TORNE `APPARATUS FOR ExTRACTING PARTICLE SIZE DATA Filed Sept. 13, 1956 May 17, 1960 G. E. GL-:Rl-umlzrr 7 Sheets-Sheet 3 May 17, 1960 G. E. GERHARDT 2,936,953

l APPARATUS FOR EXTRACTING PARTICLE SIZE DATA Filed Sept. 13, 1956 7 Sheets-Sheet 5 fion.

I l I l l SOANN/NG L/NE l TIOL`E VIDEO PUL SE SIZE RANGE I l SUBTRAGT/NG PULSE L- S/ZE RANGE GIROU/T OUTPUT r 6' INVENTOR.

GERARD ERNEST GERHARDT,

A TTORNE K May 17, 1960 G. E. GERHARDT 2,936,953

APPARATUS FOR EXTRACTING PARTICLE SIZE DATA Filed Sept. 13, 1956 7 Sheets-Sheet 6 VERY K rE v/oEo our ro SIGNAL vniega/#7:0711 00L/NTE co/vrfioL` SHAPED v/DEO D/F' f VAR/ABLE IFL/,'LOP I ,N 0L, UELAY n Nv RT -n/FF l E cf/P IN VEN TOR. GERARD ERNEST GERHARDE A TTURNE X May 17, 1960 G. E. GERHARDT' APPARATUS FOR EXTRACTING PAR'rIcu: sIzE DATA Filed sept. 13, 195s 7 Sheets-Sheet 7 Emi-22e. omkdw En INVENTOR.

GERARD E. GERHARDT ATTORNEY United States Patent O APPARATUS FOR EXTRACTING PARTICLE SIZE DATA Gerard Ernest Gerhardt, Plainleld, NJ., assignor to American Cyanamid Company, New York, N.Y., a corporation of Maine Application September 13, 1956, Serial No. 609,745 4 Claims. (Cl. A235-92) This invention relates to a device for electronically counting particles and, in a more specic aspect, to a device for counting particles above a set, predetermined size.

The counting of small particles has become a necessity in many elds of industry. Thus, for example, counts are of importance in suspension of pigments, bacteria, blood corpuscles, particles to impart soil resistance to brous materials, and the like. It is also important in many cases to determine the number of particles in certain size ranges, so-called differential counts. 'Y

In the past, particle counting has largely been done manually, forA example by counting the number of par@ ticles in a given area on a microscope slide using a suitable reticle in the microscope optics, counting particles on micrographs either from visual or electron microscopes and the like. Indirect methods have also been used, such as cent-rifuging, sedimentation and lightscatttering. These methods in some cases give reliable results, but they require a very long time; and the strain of long microscope observation or counting on personnel can be serious and, of course, as in any counting operation by human beings, the human error is always a serious possibility.

Certain proposals have been made to count particles automatically, for example the area in which the count is to be made, such as -a microscope slide, has been scanned by split beams of light from a single source which scan .the area together with two photocells, one of which can see one beam, and the other, the other. As the beams are interrupted by particles, light pulsation in the photocells results which, in the case of rapid scanning,

will normally be at video frequency. 'Ihe outputs of the two photocells are then fed into electronic circuits using gated amplifiers, the output of one beam, the guard beam or its spot of light in the scanning operation which is sometimes referred to as the guard spot, controls the gate of the amplifier, Whereas the output of the other beam constitutes the amplifier input signal. Pulse polarity is such that Ithe amplilier gate is only open so long as the guard beam encounters no particle. there will be one amplied pulse when the counting beam is interrupted by the particle at a point where the guard In other words,

beam is not so interrupted. A single pulse results for each particle and can be counted by a conventional sealer or digital computer fed from the output of the gated amplifier.

The above proposal was not practical. The adjustment of photocells and beams is extraordinarily critical and cross-talk between the two outputs is almost impossible to suppress. Severe light attenuation is an added disadvantage.

Some of the disadvantages of the multiple-photocell, multiple-beam method described above have been overcome. by using a flying spot scanning a raster on a cathode ray tube, coupled with a delay :line from the output of a single photocell equal to the period of a horizontal scanning line. The output from the delay line therefore gives signal corresponding to that whichr would have ,been

'2,936,953 Patented May 17, 1960 "ice , 2` Y produced by a guard beam and its separate photocell. This in elect splits the signal output of the photocell electronically instead of splitting the beam. The device, while more reliable, is still by no means a perfect answer, and it is with improvements to a device using a single beam and single photocell to which the present invention relates.

Neither of the proposed electronic counters describedY `above are capable of satisfactory dilferential counts, that is' to say of counting only particles above a certain size range. This problem still remains for direct solution, and the less accurate indirect methods referred to above were still needed, with all their time-consuming disadvantages' and liability to human error. The present in-V vention, in a specific aspect, solves the problem by eliminating respondence from particles having rhorizontal dimension below a predetermined minimum.

The invention will be described in greater detail in conjunction with the drawings in which:

Fig; l is a diagram of the optics and lblock diagram of the electronic circuits producing the video output;

Fig. 2 is a block diagram of the main electronic circuits;

Fig. 3 is a schematic of the wiring of the portion of Fig. 2 enclosed in the dashed rectangle; I Y

Fig. 4 is a plan view of scanning beams encountering a particle;

Fig. 5 -is a series of wave vforms observable at various numbered points on Fig. 2V for the four cases illustrated in Fig. .4;

Fig. 6 is a plan view of a particle being scanned by two beams for differential counting and the wave forms associ'ated therewith; r

Fig. 7 is a block diagram of the circuit for dilferential particle size count;

Fig. 8 is a block diagram Fig. 9 is a schematic of the resetting circuits of Fig. 2. In Fig. l there is shown a cagiode ray tube 14 of the higheintensity, shortpersistence type, such as a 5ZP16, the second anode of which is fed from va. high-voltage supply 15. Scanning of the raster on the tube 14is effected by a conventional sync generator 11 which feeds a horizontal and vertical sweep generator r13 timed by the sync generator 11 which also controls a blankingvgen'- erator 12 for eliminating retracing. The sync generators, sweep generators, blanking generator and voltage supplies to the cathode ray tube are conventional, and .these sections ofthe circuit are therefore shown diagrammatically as their particular design forms no part of the present invention.

The bright flying spot on the face of the cathode ray tube traces out a raster which is focused by the lens system 16 onto the transparent field 17 in which particles are to be counted and which in practice will usually be a microscope slide, photographic transparency or the like. As the field to'be counted is normally smaller than the raster on the cathode ray tube, the image is reduced in size, which serves .to further reduce the size of the flying spot and hence increases the precision with which particles are counted. Y

The light passing through the slide or transparency 17 is focused onto the cathode of la multiplier phototube 19 shown in diagrammatic form. The tube output from the load resistor 20 ispassed through a video preamplier 21, equalizer 22 andvideo amplifier 23. The amplifiers are The outp-ut from amplifier 423 amplifier is connected to a conventional monitoring kinescope (not shown). Likewise, there is a connection from the output of the`sync generator v11 to a delay line 10 shown in Fig. 2, this delay line in a standard circuit de# signed to introduce adelay of 63.5 as. The delay line receives pulses at horizontal sync frequency and also of a modified scanner; and,l

phase as at 3.

shaped pulses from the shaped 24, to the output of which it is connected (Fig. 2). The output point 1 of the delay line corresponds to the preceding line on the raster and acts as a guard beam or spot. At the same time there appears in the output of the shaper the output signal from the neXt scanning line, which is the active one.

The effect of scanning is shown in Figs. 4 and 5 and Will be described before continuing the description of the electronic circuits. Fig. 4 shows an enormously enlarged image of a portion of the microscope slide or transparency 17 scanned by the beam from the flying spot. Two beams are illustrated for Case I in which the upper beam is derived from the delay line l and the lower is the active scanning beam. As the delay line causes the signal from the delayed beam to appear contemporaneously with the active scanning beam, the Wave forms appear as at 1 and 2 in Fig. 5, the numerals referring yto the nurnbered points -appearing on Fig. 2. It will be seen that in VCase I the delayed beam does not encounter a particle at the instant illustrated and therefore shows no negative pulse; Whereas .the active scanning beam does encounter a particle edge, passing through a comparatively short dimension thereof, and appears at point 2 on Fig. 2 as a shaped positive square pulse of fairly short duration.

,In Case II, where both the delayed beam and the yactive beam encounter the particle, the latter encountering a larger dimension, a negative shaped pulse appears at the end of the delayline at .1; and a somewhat longer square positive pulse appears at 2. In Case III both beams encounter the particle but the dimensions are reversed and the negative pulse at l is now longer than the positive pulse at 2. Case IV deals with the situation where the delayed beam just strikes the particle, Whereas the succeeding line scanned by the active beam missesfthe particle. In this case Fig. 5 shows only the fairly short negative pulses which appears at 1.

The output of the delay line in Fig. 2 contains both signal from the phototube 19 and horizontal sync signal from the sync generator. These two -signals are then passed through a conventional sync separator circuit Where the sync pulses are separated out and fed into the sync generator l1. The delayed horizontal sync pulses equate the period of the horizontal sync oscillator to that of the delay line. This prevents drifting and assures that the delay scan and active scan are always spaced one line on the raster.

The video portion of the signal from the sync separator passes into the control input or" the first gated amplifier, the input to the amplifier at 2 receiving signal from the shaper 24; that is to say, a signal corresponding to the active scan. The signal input at 2 appears in the output of the first gated amplifier at 3 only during theperiod when there is no video pulse from the delay line caused by the scanning spot in the preceding horizontal line striking a particle. The negative pulse closes the gate in the amplifier. The output wave form 3 for the four cases to Fig. 9.

shown in Fig. 4 will be seen in Fig. 5. When the first 1 gated amplifier is open, as in Case I, the signal pulse appears as a negative pulse of the same duration as the input signal, phase reversal resulting from a single-stage gated amplifier. In the Cases III and IV, where the gated amplifier is closed for at least as long as there is anyV signal, no signal appears at 3. On the other hand, in Case II, where the signal pulse is longer than the gating pulse, two short negative pulses appear at 3.

The output from the first gated amplifier is then passed through a differentiating and a negative clipping circuit and lemerges as sharp pulses corresponding to the leadingV edge of the video pulse. The sharp pulse is in the same K The pulse corresponding 'to the trailing edge is eliminated.

The output then goes to a first fiip-fiop circuit which produces a positive square pulse at its output 5. This signal enters the second gated amplifier as control for the gate, holding it open only so long asY the positive 4 pulse continues. The signal for the second gated amplifier is a sharp spike produced from the trailing edge of the pulse from the original Shaper 24 which passes through another differentiating circuit. Wave forms at the positions 5 and 6 appear Aon Fig. 5 in Cases I and II, and the amplified Wave form at the output of the second gated amplifier at 7 likewise appears. This in turn is the signal for a third gated amplifier which is controlled by the vertical sync passing through a fourth gated amplifier and a second flip-flop circuit, the latter furnishing control for said third gated amplifier. The result of the circuit is, of course, to reset after each vertical eld. The wiring diagram for the manual resetting shown at the bottom of Fig. 2 is illustrated in Fig. 9 in which the sequence of the vertical sync input signal is shown conventionallyy from left to right. In Fig. 2, Vfor sake of symmetry, the blocks are shown with the vertical sync input signal entering at the right. The following description will refer YManual resetting is provided in conventional manner by a grounded reset button, so labeled in Figs. 2 and 9. The reset button which is a normally closed switch, as is customary, connects the grids of the second tube of the rst flip-flop, the second half of the tube of the second flip-liep and the first half of the tube of the third flip-flop through suitable resistors to ground. In its normal position, the second'half `of the tube of the third flip-flop is conducting, as is the second half of the tube of the second flip-flop. The plates being both negative, the control grids of the fourth gated amplifier and of the third gated amplifier, which feeds the sealer at 8 -in Fig. 2, are both closed. Accordingly, no signals reach the'scaler, and vertical sync input signals are not amplified. The first flip-flop between positions 4' and 5 of Figs. 2 and 3 is likewise'grounded and operates normally. When the reset button is pushed to the open position, the first half of the tube of the third fiip-op receives positive voltage from B+. This half `of the tube there fore starts to conduct, throwing a negative voltage on the grid of the second half of the tube which is cut olf. In other words, the third flip-liep is flipped. The cut off of this second half of the tube puts a positive voltage on the suppressor grid of the fourth gated amplifier and, therefore, opens this fourth amplifier gate so that vertical sync signals can now be amplied. However, as long as the reset button is held in, the grid return of the second half of the tubeVo-f the second fiip-flop` is open and, hence, the flip-flop is ineffective. As soon as the reset button is released the last-mentioned grid ref tum is closed, and the next vertical sync pulse is ampli- `fied by the fourth gated amplifier; and the positive pulse,

after passing through the irst'dicde, causes the second flip-liep to flip, the first half of the tube becoming conductive and second half of the tube cut ofi.v The plate of the latter becomes positive and imparts its positive voltage to the gate of the third gated amplifier leading to the sealer (Figs. 2 and 3). Accordingly, video pulses during the `whole of the vertical scanning field are passed through the rest of the system, as described above, and if the delay line does not signal the presence of corresponding particles in the preceding line the particles are counted by the Scaler.

When the vertical scanning field has gone through one cycle,.the next vertical sync pulse passes through the sec ond diode (Fig. 9)v in the form of a positive pulse to the second half of the tube of the second flip-nop. This -causes conduction in the tube, resetting the flip-flop to the original position, and hence, closing the gate of the third gated amplifier to the scaler. The third flipop flops `back to its original position and puts a negative voltage on the gate of the fourth gated amplifier. No further vertical sync pulses are amplified until the reset button is again pressed and released. As a result a single vertical field is scanned andthe pulses counted by the scaler. If it is desired to count more than one vertical eld,

f more flip-flops are used. z These circuits are4 conventional in. design.'v

The output at 8 from the third gated amplifier, which has;a wave form shown in Fig. 5, then passes to the input of a conventional scaler or .a digital counter which is not shown.

- It will be noted that in Case I .the pulse in the output of vthe iirst'flip-op circuit at 5 is Ilimited in duration. Otherwise, the second gated ampliiier would be kept open indefinitely until the next pulse struck the input to the flip-flop, and this would make it possibleto get a confused reading from a subsequent particle if both beams clo'se the gate in the first gated amplifier and therefore the signal coming in `at 2 would not be passed on, the secondv gated amplifier would remain open, and the signal from the lactive scan, after differentiation, would enter struck the particle because, while the delay'beam would d at.6l In other words, the subsequent particle were'-- st-ruck by both beams, it would be counted a second time. It is to prevent this inaccuracy that oneY of the important features of the present invention is provided, which feature will now be described.

A portion of the negative-going pulse from the output of the second gated amplifier at 7 is passed through a delay circuit into a buffer amplifier having one stage, the

output. being at 9. The resulting positive pulse, which is slightly delayed, is applied to the clipping circuit, where phase reversal takes place and hence a negative pulse is applied to the clipper ahead of the first flip-flop@ Thus, when the beams encounter a subsequent particle in the scan, the circuits are in preset condition, and so there be no count if both the .delay beam and .the active scanning beam encounter the particle. Within the resolution limits of the instrument, the count of each particle will always be exact, regardless of the relative positions and sizes of the particles, and a fully accurate, automatically resetting counting circuit is provided.v

Fig. 3 illustrates a typical schematic circuitl covering the portion of the block diagram in Fig. 2 enclosed with.

in the dashed lines. The first gated amplifier, -a 6AS6, is connected in conventional manner. The differentiating is accomplished by a condenser, and' resistor; clipping is done by a 6CB6; the clipped pulse- Ais amplified by a 6197; and it is to the grid of this tube that the delayed pulse isfed from the buffer amplifier, which uses a 12AU7 as amplifier `and cathode follower output.

In Fig. 3 a short delay line is shown as aphysical- However, it is sometimes not necessary to component. incorporate a special delay line as the buiferampliier introduces some delay, which in some cases is suicient to provide smooth operation. The block diagram of Fig.

2 denotes function and, of course, the delay results, re.-`

gardless of whether itis lumped or distributed. It will be referred to in the claims as a short delay circuit.

When counting particles of a predetermined particle size, the ,short delay circuit must of course., be less than that corresponding tothe time. required for the scanning spot to pass fromyone edge of the particle to the other. Otherwise, the particle will not be counted; However, as is conventional in flip-flop circuits, ythere is atinite, even though-very short, switching time in the ip-op circuits. Inyotherwords, their. response, as in practically all electronic circuits, is not instantaneous and, of course, the delay must be longer than the switch-Q ing time. As the very short switching Vtime of the flipa flop circuits will vary somewhat with the tubes and components used, no exact minimum delay time canbe given for the circuit.

The description of Figs. 1 5 has been in connection with a vraster scanning device in which a flying spot from a cathode ray tube scans the sample. This is, of course, not the only raster scanning method which can be used, and in Fig. 8 a different raster scanning method is illustrated, the same parts bearing the same reference numerals as in Fig. 1. A source of light illuminates the sample 17 which is imaged by the lens 27 onto 'the' face of a television camera tube 26, the scanning beam of which is timed by the sync generator 11 and which generates a signal output showndiagrammatically as developed across a resistance 20.y 'Ilhe signal which contains infomation at video frequency is, of course, of the same type as in Fig. l and the counting proceeds exactly ranged as the photomultiplier tube is in Fig. 1 to receive an image by transmitted light.

In the description above `a counting circuit has been described in which the delay line is equal to a single horizontal scanning period. This gives the highest resolution possible with the equipment used and may be considered the preferred embodiment of the present invention. It will be obvious, of course, that the delay may be any multiple of the horizontal scanning period, and for some purposes it is desirable to have the guard beam delayed two or more horizontal lines. The operation of the circuits is, of course, the same, regardless of what multiple of a horizontal scanning period the delay represents.

In the description above a complete raster has been described, either by a ying spot or on the face of a Atelevision camera tube. For most operations this is the preferred embodiment yofl the invention. However, if

there is a uniform ow. of liquid through a hollow cell ora uniform movement of a sample, it is possible to' scan only horizontally the movement of the' sample or flow representing the vertical scanning. In each case a raster scanning lresults, regardless of whether the sample stands still and there is a vertical displacement of the shaper 24 and the circuits in Fig. 2 to introduce a pulse' of opposite phase to the scanning pulse and of variable duration. For the duration of the size range subtracting pulse. no video signal passes to the input of Fig. 2. Fig.y

Only the portion offthe.

6 shows such an operation. particle to the right of the dotted line will produce ya scanning pulse which passes into the delay linefand the rest of the circuit; Particles which are notlarge enough to produce a scanning pulse of longer durationt-han thesubtracting pulse will not be counted. Fig. 7 illustrates in vblock diagram. the circuit for the sizelrangesubtracting pulse. vThe incoming scanning pulse is applied to two circuits, one going to a differentiating circuit, and the second through `a1 very short delay line Vto an inverting circuit Kand then to a differentiating.

circuit. The very short .delay line has a delay which is shorter than the time taken by the scanning beam to pass over the smallest sizeof particle which the instrument is capable of counting. This very short delay line, however, must introduce a finite delay at least equal to the switching time of the fourth dip-flop circuit described below. Each differentiating circuit includes a negative clipping diode. From the rst differentiating circuit the positive pulse corresponding to the leading edge is fed directly to one tube of a flip-flop circuit and also to a variable delay line which is, of course, shorter than the maindelay line separating delay beam from scanning beam.- The output of this variable delay line and the.

Vpassed on tothe circuits of Fig. 2.

positive pulse from the trailing edge Iin the second differ; circuit are passed to the other tube. of the. iiiptionk of the video pulse is longer than variable delay, the

iifth gated amplifier is opened while there is still a video signal on the input thereof. Video signal is therefore If the variable delay is longer than the video pulse, there will be no rvideo on the fifth gated ampliiier input when it is opened. As a result particles smaller than determined by the variable delay will not be counted.

In differential scanning, of course, the variable delay is set for a certain minimum particle size. A count is then made which will only count particles largervthan thispredetermined minimum. The circuit may be used to make a number of counts of different particle sizes by successive settings of the variable delay line so that a complete size range story can be obtained.

I claim:

comprising a differentiating` and clipping circuitand vari:-`

able delay means, an additional circuit from the input;

tothe gatedr amplifier tothe output of the variable delay means, said circuit comprising inverting, clipping. and differentiating means, the. clipping being of a polarity to produce differentiated pulses of the same. polarity as thosein` the output ofthe variable delay means whereby the. gated amplifier is closed with the leading edge of a video `frequency signal pulse and is not opened until a pulse from the variable delay means or the inverting, clipping and' differentiating circuit actuates the gate..

3. A device according to claim 2 in which the gate actuating circuit is a ip-op.

4. In a particle counting device for counting particles above a predetermined size comprising a holder adapted to hold a sample in a predetermined plane, raster scanning meansoptically associated with said plane, a photof l. In a particle` counting device for counting particles Y above a predetermined size comprising a holder adapted to hold a sample in a predetermined plane, raster scan ningmeans optically associated with said plane, a photoelectric detector for producing electric pulses at video4 frequency as discreteV particles in the plane are scanned, anelectronic` counter therefor including a main delay circuit! having a delay periodV equal to a multiple of a horizontal scanning period, feedback means from said mainl delay circuit to the raster scanning means to insureV synchronous operation, and triggered gating means actuated by the output of the delay circuit to form a coincidence-anticoincidence circuit, counting taking place only n Whenno pulse from the delay line output encounters the.

triggered gating means, the improvement which comprises a short delay circuit having a delay less than the time taken by the scanning means to pass over the smallest particle size to be counted but greater than the switching time of. the triggered gating means and means for connecting a portion of a signal from said detector to the input ofy said circuit and to connect in proper phase the output of said circuit Vto the triggered gating. means.

2. A device according to claim l comprising circuits inthe input of the main delay circuit to transmit signals from predetermined particle size ranges, said vcircuits comprising a gated ,ampliiien the output of which is connected` to the input of the delay circuit, a veryshort delay circuit in the signal input of the gated ampliiier, av control means actuating the amplier gate, connecting means from the video frequency pulse output to the very short delay line having a delay. less` than the time required by the scanning means topass over the smallest particle size to be counted. but greater than the switching time ofthe ampliiier `gate actuating means and to the gate actuating circuit in phase tc close the gate, a circuit to the gate control circuit to open the gate, said circuit Velectric detector. for producing electric pulses at video frequency as discrete particles in the plane are scanned,

. an electronic counter therefor includingadelay circuit having a delay period equal to a multiple of a horizontal scanning period and triggered gating means. actuated byv `the output of the delay circuit to form a coincidenceanticolncidence circuit, counting taking place only when noA pulse from the delay circuit output encountersthe triggered gating means, the improvement which comprises` circuits in theinput of the delay circuit, said circuits comprisinga gated amplifier, the output of which is connected to the input of the `delay circuit, averyl short delay circuit in the signal input of the gated amplifier,- a control means actuating the amplifier gate the delay ofl the very short delay circuit being lessthan the time.

requiredtby the scanning means to pass over the smallest particle size to be counted but greater than the switchingk time of the ampliiier gate actuating means, connecting means from the input to the very short delay circuit'to..

thegate actuating circuit in phase-to close the gate,

a shunt circuit to a portion of the gate control circuitl to open the gate, said shunt circuit comprising a variable delay line, a second shunt circuit from the input to the gated ampliiier to the output of the variable delay means, said circuit comprising inverting, clipping and differentiating means, the clipping beingoffa polarity to produce diiferentiated pulses of the same polarity as those in the output of the variable delay means whereby the gated ampliiier is closed with the leading edge ofa signal pulse and is not opened until a pulse from the variable delay means or the inverting, diierentiating and-clipping circuit actuates the gate.

British Journal of Applied Physics Supplement No. 3,

April 1954, Par. 1, page S123, Section 8, page S124, andA pages S156 rto S159. i v 

